Stable parenteral DNJ compositions

ABSTRACT

A stable pharmaceutical composition that includes an active agent selected from 1-deoxynojirimycin, a pharmaceutically acceptable salt thereof, or a derivative thereof, and a buffer, wherein the stable pharmaceutical composition is capable of being parenterally administered to a human without deleterious health effects. Pompe disease is an example of a lysosomal storage disorder. Pompe disease is caused by a deficiency in the enzyme acid alpha-glucosidase (GAA). GAA metabolizes glycogen, a storage form of sugar used for energy, into glucose.

REFERENCE TO RELATED APPLICATIONS

The present application is a continuation of U.S. application Ser. No.14/759,770, which is the National Phase entry of PCT/US14/10891, filedJan. 9, 2014, claims the benefit of U.S. Provisional Application No.61/750,677, filed Jan. 9, 2013, and U.S. Provisional Application No.61/914,839, filed Dec. 11, 2013, each of which being hereby incorporatedby reference in their entirety.

BACKGROUND

Pompe disease is an example of a lysosomal storage disorder. Pompedisease is caused by a deficiency in the enzyme acid alpha-glucosidase(GAA). GAA metabolizes glycogen, a storage form of sugar used forenergy, into glucose. The accumulation of glycogen is thought to lead toprogressive muscle myopathy throughout the body which affects variousbody tissues, particularly the heart, skeletal muscles, liver, andnervous system. According to the National Institute of NeurologicalDisorders and Stroke, Pompe disease is estimated to occur in about 1 in40,000 births.

There are three recognized types of Pompe disease—infantile, juvenile,and adult onset. Infantile is the most severe, and presents withsymptoms that include severe lack of muscle tone, weakness, enlargedliver and heart, and cardiomyopathy. Swallowing may become difficult andthe tongue may protrude and become enlarged. Most children die fromrespiratory or cardiac complications before the age of two. Juvenileonset Pompe disease first presents in early to late childhood andincludes progressive weakness of the respiratory muscles in the trunk,diaphragm, and lower limbs, as well as exercise intolerance. Mostjuvenile onset Pompe patients do not live beyond the second or thirddecade of life. Adult onset symptoms involve generalized muscle weaknessand wasting of respiratory muscles in the trunk, lower limbs, anddiaphragm. Some adult patients are devoid of major symptoms or motorlimitations.

Enzyme replacement therapy (ERT) is one approach to treating PompeDisease. One of the main complications with ERT is the attainment andmaintenance of therapeutically effective amounts of enzyme due to rapiddegradation of the infused enzyme. As a result, ERT requires numerous,high-dose infusions and is costly and time consuming.

A newer approach to treating Pompe Disease, a specific chaperonestrategy, rescues mutated proteins from degradation presumably in theendoplasmic reticulum (ER) or in other cellular proteindegradation/disposal systems. This strategy employs small moleculereversible inhibitors which specifically bind to a defective lysosomalenzyme associated with a particular lysosomal disorder. The chaperonestrategy involves the use of a small molecule that facilitates thecorrect folding of a mutated protein, to prevent undue or abnormaldegradation from the ER quality control system. These specificchaperones are designated as active site-specific chaperones. See, e.g.,U.S. Pat. Nos. 6,274,597, 6,583,158, 6,589,964, and 6,599,919, each ofwhich incorporated by reference herein. U.S. Pat. No. 6,583,158discloses 1-deoxynojirimycin (DNJ) and other active site-specificchaperones for treating Pompe Disease.

1-DNJ can be provided in a solid form for oral administration. However,if DNJ is to be administered as part of a combination therapy along withERT, there is a desire for parenterally administered DNJ, as aparenteral formulation allows for greater dosage control andadministration access. Also, patients, expecially infants, may havedifficulty swallowing oral dosage forms, regardless of whethermonotherapy or combination therapy is elected as a form of treatment.Although 1-DNJ is soluble and generally stable in aqueous formulations,such formulations are not suited for parenteral administration becauseof the need for pH control over a practical shelf-life of theformulation and to provide a suitable osmolality for parenteraladministration such that the formulation does not cause deleterioushealth effects.

BRIEF SUMMARY OF THE INVENTION

One aspect of the presently disclosed subject matter provides a stablepharmaceutical composition that includes an active agent selected from1-deoxynojirimycin, a pharmaceutically acceptable salt thereof, or aderivative thereof, and a buffer, wherein the stable pharmaceuticalcomposition is capable of being parenterally administered to a humanwithout deleterious health effects.

The active agent can be, for example, 1-deoxynojirimycin hydrochlorideor N-butyl-deoxynojirimycin, or a pharmaceutically acceptable saltthereof (e.g., N-butyl-deoxynojirimycin). In certain embodiments, thebuffer can be selected from a phosphate buffer (e.g., a sodium phosphatebuffer), a citrate buffer (e.g., a sodium citrate buffer), an acetatebuffer (e.g., sodium acetate buffer) and a bicarbonate buffer (e.g., asodium bicarbonate buffer). The pH of the pharmaceutical composition canbe from about 4.6 to about 5.5 (e.g., 5.0). The pharmaceuticalcomposition can be formulated for subcutaneous administration, orintravenous administration. In certain embodiments, the pharmaceuticalcomposition demonstrate a lack of hemolytic potential.

In certain embodiments, the active agent (e.g., 1-DNJ-HCl) can bepresent at a concentration of from about 25 mg/mL to about 30 mg/mL andthe buffer is present at a concentration of from about 40 mM to about 50mM. In certain embodiments, the active agent (e.g., 1-DNJ) can bepresent at a concentration of about 25 mg/mL or about 30 mg/mL and thebuffer is present at a concentration of about 40 mM or 50 mM. The totalamount of active agent can be, for example, 300 mg and the total volumeof the composition can be, for example, about 10.5 mL.

In certain embodiments, the pharmaceutical composition can furtherinclude a chelating agent (e.g., EDTA). For example, the active agentcan be present at a concentration of about 30 mg/mL, the buffer can bepresent at a concentration of about 50 mM, and EDTA can be present at aconcentration of about 0.05% weight by volume. In one embodiment, theactive agent can be present at a concentration of about 25 mg/mL, thebuffer can be present at a concentration of about 50 mM, and EDTA can bepresent at a concentration of about 0.05% weight by volume.

In certain embodiments, the composition can be shelf-stable for at leastabout one year under a nitrogen atmosphere and a temperature of fromabout 2° C. to about 42° C. In one embodiment, the composition can beshelf-stable for at least about one year under a nitrogen atmosphere anda temperature of from about 2° C. to about 8° C. In another embodiment,the composition can be shelf-stable for at least about one year under anitrogen atmosphere and a temperature of from about 23° C. to about 27°C. In another embodiment, the composition can be shelf-stable for atleast about one year under a nitrogen atmosphere and a temperature offrom about 38° C. to about 42° C.

In one embodiment, the composition can be shelf-stable for at leastabout one year under the air atmosphere or a combination of air andnitrogen atmosphere and a temperature of from about 2° C. to about 8° C.In another embodiment, the composition can be shelf-stable for at leastabout one year under the air atmosphere or a combination of air andnitrogen atmosphere and a temperature of from about 23° C. to about 27°C. In another embodiment, the composition can be shelf-stable for atleast about one year under the air atmosphere or a combination of airand nitrogen atmosphere and a temperature of from about 38° C. to about42° C.

A second aspect of the presently disclosed subject matter provides amethod for increasing the stability of an active agent selected from1-deoxynojirimycin, a pharmaceutically acceptable salt thereof, or aderivative thereof in a formulation for a parenteral administration to ahuman, that includes introducing a buffer to the active agent to preparethe formulation and storing the formulation under the nitrogenatmosphere or under the air atmosphere or a combination of air andnitrogen atmosphere and a temperature from about 2° C. to about 42° C.In some embodiments the atmosphere is a nitrogen atmosphere. In someembodiments the temperature is below room temperature (e.g., from about2° C. to about 8° C.). In some embodiments the temperature is from about23° C. to about 27° C. In some embodiments the temperature is from about38° C. to about 42° C. The formulation can be shelf-stable for at leastone year, or at least two years.

Another aspect of the presently disclosed subject matter provides a kitfor parenteral administration to a human of a stable pharmaceuticalcomposition that includes an active agent selected from1-deoxynojirimycin, a pharmaceutically acceptable salt thereof, or aderivative thereof, and a buffer, wherein the composition isshelf-stable for at least about one year under a nitrogen atmosphere orshelf-stable for at least about one year under an air atmosphere orshelf-stable for at least about one year under a combination of air andnitrogen atmosphere and a temperature of from about 2° C. to about 42°C. In some embodiments the atmosphere, for which the formulation isstable, is a nitrogen atmosphere. In some embodiments the temperature,for which the formulation is stable is below room temperature (e.g.,from about 2° C. to about 8° C.). In some embodiments the temperature,for which the formulation is stable, is from about 23° C. to about 27°C. In some embodiments the temperature, for which the formulation isstable, is from about 38° C. to about 42° C.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows stability of 1-deoxynojirimycin hydrochloride (1-DNJ-HCl orAT2220) in sodium phospahate buffer solution compared to stability inwater as described in Example 2. The stability was measured by normalphase high performance liquid chromatography (NP-HPLC). The left spectrashows 1-deoxynojirimycin as detected by UV detector, while the rightspectra show 1-deoxynojirimycin as detected by Charged Aerosol Detector(CAD).

FIG. 2 shows stability of 1-deoxynojirimycin chloride in sodium acetatebuffer solution compared to stability in water as described in Example3. The stability was measured by normal phase high performance liquidchromatography (NP-HPLC). The left spectra shows 1-deoxynojirimycin asdetected by UV detector, while the right spectra show 1-deoxynojirimycinas detected by Charged Aerosol Detector (CAD).

FIG. 3 shows stability of 1-deoxynojirimycin chloride in sodium citratebuffer solution with an addition of EDTA under the nitrogen atmosphereas described in Example 4. The stability was measured by normal phasehigh performance liquid chromatography (NP-HPLC). The left spectra shows1-deoxynojirimycin as detected by UV detector, while the right spectrashow 1-deoxynojirimycin as detected by Charged Aerosol Detector (CAD).

FIG. 4 shows stability of 1-deoxynojirimycin chloride in sodium citratebuffer solution compared to stability in water as described in Example5. The stability was measured by normal phase high performance liquidchromatography (NP-HPLC). The left spectra shows 1-deoxynojirimycin asdetected by UV detector, while the right spectra show 1-deoxynojirimycinas detected by Charged Aerosol Detector (CAD).

FIG. 5 shows stability of 1-deoxynojirimycin chloride in sodium citratebuffer solution compared to stability in water as described in Example6. The stability was measured by normal phase high performance liquidchromatography (NP-HPLC). The left spectra shows 1-deoxynojirimycin asdetected by UV detector, while the right spectra show 1-deoxynojirimycinas detected by Charged Aerosol Detector (CAD).

DETAILED DESCRIPTION OF THE INVENTION

The presently disclosed parenteral formulations include 1-DNJ (alsoreferred to as DNJ), or a salt or derivative thereof. In one embodiment,the parenteral formulation includes 1-DNJ-HCl. In another embodiment,the formulation includes a derivative of 1-DNJ, such as N-butyldeoxynojirimycin. The derivative of 1-DNJ (e.g., N-butyldeoxynojirimycin) may be found in addition to, or in place of, 1-DNJ or1-DNJ-HCl.

As noted above, the active agent is selected from 1-deoxynojirimycin, apharmaceutically acceptable salt thereof, or a derivative thereof. Theconcentration of the active agent in the formulation can, in certainnon-limiting embodiments, range from about 10 mg/ml to about 100 mg/ml,or from about 20 mg/mgl to about 50 mg/ml, or from about 20 mg/ml toabout 35 mg/ml. In some embodiments the concentration of the activeagent in the formulation can be 20 mg/ml, 21 mg/ml, 22 mg/ml, 23 mg/ml,24 mg/ml, 25 mg/ml, 26 mg/ml, 27 mg/ml, 28 mg/ml, 29 mg/ml, 30 mg/ml, 31mg/ml, 32 mg/ml, 33 mg/ml, 34 mg/ml or 35 mg/ml. In some embodiments theconcentration of the active agent in the formulation can be about 30mg/ml. In some embodiments concentration of 1-deoxynojirimycin as a freebase in the formulation can be about 25 mg/ml

In certain embodiments, the pH of the formulation is from about 3 toabout 9 or to about 10, or from about 3 to about 6, or from about 3.0 toabout 5.5. In one embodiment, the pH of the formulation is below 6.0, orbelow 5.5. It has been found that discoloration may occur in1-DNJ/buffer formulations with a pH in the range of about 6.0-7.5. Insome embodiments the pH of the formulation can be 3.0, 3.1, 3.2, 3.3,3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7,4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9 or 6.0. Insome embodiments the pH of the formulation can be about 5.0.

The formulation can contain a buffer. In one embodiment, the buffer isselected from sodium citrate, sodium acetate and sodium phosphate. Inone embodiment, the buffer is sodium citrate. Other salts known to beused with buffers, besides sodium salts, can also be used, such aspotassium and ammonium salts. Thus, the buffer can be, for example, apotassium citrate, potassium acetate or potassium phosphate buffer, oran ammonium citrate, ammonium acetate or ammonium phosphate buffer. Theconcentration of the buffer can, in certain non-limiting embodiments,range from about 20 mM to about 75 mM, or from about 30 mM to about 60mM, or from about 35 mM to about 55 mM. In some embodiments theconcentration of the buffer can be 30 mM, 31 mM, 32 mM, 32 mM, 33 mM, 34mM, 35 mM, 36 mM, 37 mM, 38 mM, 39 mM, 40 mM, 41 mM, 42 mM, 45 mM, 46mM, 47 mM, 48 mM, 49 mM, 50 mM, 51 mM, 52 mM, 53 mM, 54 mM, 55 mM, 56mM, 57 mM, 58 mM, 59 mM or 60 mM. In some embodiments the concentrationof the buffer can be about 50 mM.

In some embodiments the formulation includes a chelating agent, such asEDTA or EGTA. In some embodiments, EDTA is present in the formulation ata concentration from 0.005% to 0.25% (w/v), or from 0.01% to 0.1% (w/v).In one embodiment, EDTA is present in the formulation at a concentrationof about 0.05% (w/v).

In one embodiment, the presently disclosed parenteral formulation isstored under a nitrogen atmosphere. In one embodiment, the presentlydisclosed parenteral formulation is stored under an air atmosphere. Inone embodiment, the presently disclosed parenteral formulation arestored under a combination of nitrogen and air atmosphere. In oneembodiment, the presently disclosed parenteral formulation are stored atbelow 45° C., or below 25° C., or below 15° C., or below 10° C.

In one embodiment, the osmolality of the presently disclosed parenteralformulation ranges from about 280 to about 320 mOsm/kg. In analternative embodiment, the osmolality of the presently disclosed isbelow about 500 mOsm/kg, or below about 450 mOsm/kg. In one embodiment,the concentration of 1-DNJ-HCl is about 30 mg/mL, the concentration ofthe buffer is 40 mM and osmolality of about 374 mOsm/kg. In anotherembodiment, the concentration of 1-DNJ as a free base is about 25 mg/mL,the concentration of the buffer is 40 mM and osmolality of about 374mOsm/kg. In another embodiment, the concentration of 1-DNJ-HCl is about30 mg/mL, the concentration of the buffer is 50 mM and osmolality ofabout 400 mOsm/kg. In another embodiment, the concentration of 1-DNJ asa free base is about 25 mg/mL, the concentration of the buffer is 50 mMand osmolality of about 400 mOsm/kg. In other embodiments 1-DNJ-HClconcentration will be from about 30 mg/mL to about 60 mg/mL.

In one embodiment the formulation is suitable for subcutaneousadministration. In one embodiment the formulation is suitable forintravenous (IV) administration.

In one embodiment, the 1-DNJ-HCl is present at a concentration of about30 mg/mL and the buffer is present at a concentration of about 50 mM atpH of about 5. In one embodiment, the 1-DNJ, as a free base is presentat a concentration of about 25 mg/mL and the buffer is present at aconcentration of about 50 mM at pH of about 5. In one embodiment, theformulation is stored under the nitrogen atmosphere and at a temperatureof from about 2° C. to about 8° C. In one embodiment, the formulation isstored under the nitrogen atmosphere and at a temperature of from about23° C. to about 27° C. In one embodiment, the formulation is storedunder the nitrogen atmosphere and at a temperature of from about 38° C.to about 42° C.

In one embodiment, the pharmaceutical composition contains 30 mg/mL of1-DNJ-HCL at pH 5 and a 40 mM sodium citrate buffer. In one embodiment,the pharmaceutical composition contains 25 mg/mL of 1-DNJ, as a freebase at pH 5 and a 40 mM sodium citrate buffer. In one embodiment, thepharmaceutical composition contains 30 mg/mL of 1-DNJ-HCL at pH 5, 40 nMsodium citrate buffer and 0.05% w/v EDTA. In one embodiment, thepharmaceutical composition contains 25 mg/mL of 1-DNJ, as a free base atpH 5, 40 nM sodium citrate buffer and 0.05% w/v EDTA. In one embodiment,the pharmaceutical composition contains 30 mg/mL of 1-DNJ-HCL at pH 5and a 40 mM sodium acetate buffer. In one embodiment, the pharmaceuticalcomposition contains 25 mg/mL of 1-DNJ, as a free base, at pH 5 and a 40mM sodium acetate buffer. In one embodiment, the pharmaceuticalcomposition contains 30 mg/mL of 1-DNJ-HCL at pH 5, 40 mM sodium acetatebuffer and 0.05% w/v EDTA. In one embodiment, the pharmaceuticalcomposition contains 25 mg/mL of 1-DNJ, as a free base, at pH 5, 40 mMsodium acetate buffer and 0.05% w/v EDTA.

In one embodiment, the pharmaceutical composition contains 30 mg/mL of1-DNJ-HCL at pH 5 and a 50 mM sodium citrate buffer. In one embodiment,the pharmaceutical composition contains 30 mg/mL of 1-DNJ-HCL at pH 5,50 nM sodium citrate buffer and 0.05% w/v EDTA. In one embodiment, thepharmaceutical composition contains 30 mg/mL of 1-DNJ-HCL at pH 5 and a50 mM sodium acetate buffer. In one embodiment, the pharmaceuticalcomposition contains 30 mg/mL of 1-DNJ-HCL at pH 5, 50 mM sodium acetatebuffer and 0.05% w/v EDTA.

In one embodiment, the pharmaceutical composition contains 25 mg/mL of1-DNJ, as a free base, at pH 5 and a 50 mM sodium citrate buffer. In oneembodiment, the pharmaceutical composition contains 25 mg/mL of 1-DNJ,as a free base, at pH 5, 50 nM sodium citrate buffer and 0.05% w/v EDTA.In one embodiment, the pharmaceutical composition contains 25 mg/mL of1-DNJ, as a free base, at pH 5 and a 50 mM sodium acetate buffer. In oneembodiment, the pharmaceutical composition contains 25 mg/mL of 1-DNJ,as a free base, at pH 5, 50 mM sodium acetate buffer and 0.05% w/v EDTA.

Other pharmaceutically acceptable excipients which may be included inthe formulation as buffers include bicarbonate buffer, amino acids,urea, alcohols, ascorbic acid, phospholipids; proteins, such as serumalbumin, collagen, and gelatin; salts such as EDTA or EGTA, and sodiumchloride; liposomes; polyvinylpyrollidone; sugars, such as dextran,mannitol, sorbitol, and glycerol; propylene glycol and polyethyleneglycol (e.g., PEG-4000, PEG-6000); glycerol; glycine or other aminoacids; and lipids. A non-exclusive list of acceptable excipients islisted in the Table 1.

TABLE 1 List of excipients Excipient pH Range Acetate Sodium 2.5-7.0Acetic acid 2.5-7.2 Glacial acetic acid 3.5-7.0 Ammonium 6.8-7.8Ammonium sulfate — Ammonium hydroxide — Arginine 7.0-7.4 Aspartic acid5.0-5.6 Benzene sulfonic acid 3.25-3.65 Benzoate sodium/acid 3.5-6.9Bicarbonate, sodium  5.5-11.0 Boric acid/sodium — Carbonate, sodium 4.0-11.0 Citrate Acid 2.5-9.0 Sodium 3.0-8.5 Disodium 6.1 Trisodium 6.1Diethanolamine  9.5-10.5 Glucono delta lactone 5.5-7.0 Glycine/glycineHCl  2.5-10.8 Histidine/histidine HCl 5.0-6.5 Hydrochloric acid Broadrange Hydrobromic acid 3.5-6.5 Lactate sodium/acid 2.7-5.8 Lysine (L) —Maleic acid 3.0-5.0 Megulmine  6.5-11.0 Methanesulfonic acid 3.2-4.0Monoethanolamine 8.0-9.0 Phosphate Acid 6.5-8.5 Monobasic potassium6.7-7.3 Dibasic potassium 6.7-7.3 Monobasic sodium 2.5-8.0 Dibasicsodium 2.5-8.3 Tribasic sodium — Sodium hydroxide Broad range Succinatesodium/disodium 5.0-6.0 Sulfuric acid 3.0-7.0 Tartrate sodium/acid2.5-6.2 Tromethamine (Tris) 6.5-9.0Adapted from: Nema, S et. al. (2011). Excipients and their role inapproved injectable products: current usage and future directions. PDA JPharm Sci and Tech, 65, 287-332.

EXAMPLES Example 1. Preparation of Buffers

The following representative buffer compositions were prepared byadmixing the ingredients set forth below. The buffers used in Examples2-6 below were prepared in a similar fashion, amounts of acidic andbasic components modified, if necessary, to obtain the pH parametersspecified therein.

TABLE 2 Components of 50 mM sodium citrate buffer, pH 5 Component Amountper ml Sodium Citrate Dihydrate 9.32 mg Citric Acid Monohydrate 3.85 mgWater q.s. to 1.0 mL

TABLE 3 Components of 50 mM sodium acetate buffer, pH 5.5 ComponentAmount per ml Sodium Acetate Trihydrate 5.92 mg Glacial Acetic Acid0.384 mg Water q.s. to 1.0 mL

TABLE 4 Components of 100 mM sodium phosphate buffer, pH 7 ComponentAmount per ml Sodium Phosphate Monobasic Monohydrate 4.79 mg SodiumPhosphate Dibasic, Anhydrous 9.26 mg Water q.s. to 1.0 mL

Deionized and/or distilled water can be used in place of water.

Example 2. Stability and Integrity of 1-Deoxynojirimycin Chloride in 100mM Sodium Phosphate Buffer Solution

1-DNJ-HCl was dissolved in sodium phosphate buffer. 1-DNJ-HClconcentration was 60 mg/mL; sodium phosphate buffer concentration was100 mM at pH 4.89. The formulation was stored at 60° C. for 4 days.1-DNJ-HCl solution in water was used as a control. Reaction rate forsmall molecules such as 1-DNJ-HCl generally follows Arrhenius equationwhere reaction rate nearly doubles for every 10° C. increase in thetemperature. Thus, under this model, the reaction rate at 60° C. wouldbe 2⁵ or 32 times the reaction rate at 10° C. Therefore during 4 days at60° C. 1-DNJ-HCl would degrade by approximately the same percentage asduring 128 days (4*32) at 10° C.

The stability of 1-DNJ-HCl was measured by normal phase high performanceliquid chromatography (NP-HPLC). 1-DNJ-HCl was recorded by UVchromatograms and by Charged Aerosol Detector (CAD) chromatograms.Mobile phase consisted of ACN/30 mM Ammonium Acetate (90/10, v/v). Theflow rate was 1.0 ml/min with 5 μl injection at 400° C. UV absorbancewas measured at 220 nm. CAD detector settings were 35.0 psi, Gain: 100pA.

Results:

The results are shown in the FIG. 1. 1-DNJ-HCl was still present in thesodium phosphate buffer solution after 4 days at 60° C. Additionally,both UV and CAD chromatograms detected presence of new compounds in thephosphate buffer solution. The peaks, corresponding to the new compoundsare highlighted by the red arrows on the FIG. 1. Additionally adiscoloration of the phosphate buffer solution was observed. No newcompounds were detected in the control 1-DNJ-HCl aqueous solution. Theaqueous solution remainded clear after 4 days at 60° C.

Inclusion of 1-DNJ-HCl in a sodium phosphate buffer in this Exampleresulted in discoloration of 1-DNJ-HCl. Furthermore, High PerformanceLiquid Chromatography (HPLC) indicated presence of additional compoundsin 1-DNJ-HCl formulation in the sodium phosphate buffer. The presence ofadditional compounds indicated that 1-DNJ-HCl is not stable in thesodium phosphate buffer under the conditions of this Example.

Example 3. Stability and Integrity of 1-Deoxynojirimycin Chloride in 50mM Sodium Acetate Buffer Solution

1-DNJ-HCl was dissolved in sodium acetate buffer. The 1-DNJ-HClconcentration was 60 mg/mL; sodium acetate buffer concentration was 50mM at pH 5.5. The formulation was stored at 60° C. for 4 days. 1-DNJ-HClsolution in water was used as control. Reaction rate for small moleculessuch as 1-DNJ-HCl generally follows the Arrhenius equation, in which thereaction rate nearly doubles for every 10° K increase in thetemperature. Thus the reaction rate, under this model, at 60° C. wouldbe 2⁵ or 32 times the reaction rate at 10° C. Therefore during 4 days at60° C. 1-DNJ-HCl would degrade by approximately same percentage asduring 128 clays at at 10° C.

The stability of 1-DNJ-HCl was measured by NP-HPLC. Presence of1-DNJ-HCl was recorded by UV absorbance detector and by Charged AerosolDetector. Mobile phase consisted of ACN/30 mM Ammonium Acetate (90/10,v/v). The flow rate was 1.0 ml/min with 5 μl injection at 400 C. UVabsorbance was measured at 220 nm. CAD detector settings were 35.0 psi,Gain: 100 pA.

Results:

The results are shown in the FIG. 2. 1-DNJ-HCl was still present in thesodium acetate buffer solution after 4 days at 60° C. Additionally, bothUV and CAD chromatograms detected presence of new compounds in theacetate buffer solution. The peaks, corresponding to the new compoundsare highlighted by the red arrows in FIG. 2. Additionally adiscoloration of the acetate buffer solution was observed. No newcompounds were detected in the control 1-DNJ-HCl aqueous solution. Theaqueous solution remainded clear after 4 days at 60° C.

Example 4. Stability and Integrity of 1-Deoxynojirimycin Chloride in 50mM Sodium Citrate Buffer Solution with EDTA Under Nitrogen Atmosphere

1-DNJ-HCl was dissolved in sodium citrate buffer that further includesEDTA. 1-DNJ-HCl concentration was 60 mg/mL; sodium citrate bufferconcentration was 50 mM at pH 4.6. The EDTA concentration was 0.05% w/v.Some samples were stored under the nitrogen atmosphere (essentially 100%nitrogen) while others were stored under air. The formulation was storedat 60° C. for 8 days.

Stability of 1-DNJ-HCl was measured by NP-HPLC. Presence of 1-DNJ-HClwas recorded by UV absorbance detector and by Charged Aerosol Detector.Mobile phase consisted of ACN/30 mM Ammonium Acetate (90/10, v/v). Theflow rate was 1.0 ml/min with 5 μl injection at 400 C. UV absorbance wasmeasured at 220 nm. CAD detector settings were 35.0 psi, Gain: 100 pA.

Results:

The results are shown in the FIG. 3. 1-DNJ-HCl was still present in thesodium citrate buffer solution after 8 days at 60° C. No new compoundswere detected in the samples stored under nitrogen atmosphere. Thesolution remained clear with no visible discoloration. A combination ofnitrogen atmosphere and EDTA preserved stability and integrity ofAT2220. The samples stored under air showed presence of new compounds inthe acetate buffer solution. The amount of these compounds was lowerthan in Examples 1 and 2, based on number and size of the additionalpeaks. The peaks corresponding to the new compounds are highlighted bythe red arrows in FIG. 3. Additionally slight discoloration of thecitrate buffer formulation, stored under air was observed.

Example 5. Stability and Integrity of 1-Deoxynojirimycin Chloride in 50mM Sodium Citrate Buffer Solution

1-DNJ-HCl was dissolved in sodium citrate buffer. 1-DNJ-HClconcentration was 60 mg/mL; sodium citrate buffer concentration was 50mM at pH 4.7. The formulation was stored at 60° C. for 8 days. 1-DNJ-HClsolution in water was used as control.

Stability of 1-DNJ-HCl was measured by NP-HPLC. Presence of 1-DNJ-HClwas recorded by UV absorbance detector and by Charged Aerosol Detector.Mobile phase consisted of ACN/30 mM Ammonium Acetate (90/10, v/v). Theflow rate was 1.0 ml/min with 5 μl injection at 400 C. UV absorbance wasmeasured at 220 nm. CAD detector settings were 35.0 psi, Gain: 100 pA.

Results:

The results are shown in the FIG. 4. 1-DNJ-HCl was still present in thesodium citrate buffer solution after 8 days at 60° C. Based on theseresults, EDTA does not appear required for 1-DNJ-HCl formulationstability. No new compounds were detected in the control 1-DNJ-HClaqueous solution. No discoloration was observed in the aqueous solutionafter 8 days at 60° C.

Example 6. Stability and Integrity of 1-Deoxynojirimycin Chloride in 50mM Sodium Citrate Buffer Solution Under Nitrogen Atmosphere

1-DNJ-HCl was dissolved in sodium citrate buffer. 1-DNJ-HClconcentration was 60 mg/mL; sodium citrate buffer concentration was 50mM at pH 5.0. The formulation was stored at 60° C. for 17 days. Thesamples were stored under the nitrogen atmosphere (essentially 100%nitrogen). 1-DNJ-HCl solution in water was used as control. Reactionrate for small molecules such as 1-DNJ-HCl generally follows Arrheniusequation where reaction rate nearly doubles for every 10° K increase inthe temperature. Thus reaction rate at 60° C. would be 2⁵ or 32 timesthe reaction rate at 10° C. Therefore during 17 days at 60° C. AT2220would degrade, under this model, by approximately same percentage asduring 17 multiplied by 32 days or about 17 months.

Stability of 1-DNJ-HCl was measured by NP-HPLC. Presence of 1-DNJ-HClwas recorded by UV absorbance detector and by Charged Aerosol Detector.Mobile phase consisted of ACN/30 mM Ammonium Acetate (90/10, v/v). Theflow rate was 1.0 ml/min with 5 μl injection at 400° C. UV absorbancewas measured at 220 nm. CAD detector settings were 35.0 psi, Gain: 100pA.

Results:

The results are shown in FIG. 4. 1-DNJ-HCl was still present in thesodium citrate buffer solution after 17 days at 60° C. under nitrogenatmosphere. No new compounds were detected in the control 1-DNJ-HClaqueous solution. No discoloration was observed in the aqueous solutionafter 17 days at 60° C.

In summary, 1-DNJ-HCl formulations in sodium phosphate and sodiumacetate buffers resulted in discoloration. Furthermore, HPLC indicatedpresence of additional compounds in 1-DNJ-HCl formulation in the sodiumphosphate and sodium acetate buffer. In contrast, 1-DNJ-HCl formulationin sodium citrate buffer showed little to no discoloration and little tono presence of additional compounds. Based on the results of theseExamples, 1-DNJ-HCl formulation is most stable in sodium citrate buffer.

Storage of samples under nitrogen atmosphere (essentially 100%) nitrogenpreserves stability and integrity of 1-deoxynojirimycin chlorideformulation greater than storage under air. (See examples 3 and 5).

Based on the results of these Examples, the presence of EDTA is notnecessary for preserving stability and integrity of 1-deoxynojirimycinchloride formulations. (See examples 4 and 5).

Example 7. Measurement of 1-Deoxynojirimycin Chloride Osmolality inSodium Citrate Buffer

The molallity of 1-deoxynojirimycin chloride (1-DNJ-HCl) was determinedby freezing point depreciation. 1-deoxynojirimycin chlorideconcentration ranged from 30 mg/mL to 60 mg/mL. Sodium citrateconcentration ranged from 20 to 50 mM.

Results:

The results are presented below in Table 5.

TABLE 5 1-deoxynojirimycin chloride osmolality Osmolality (mOsm/kg)1-DNJ- Sodium Citrate Buffer Concentration HCl 50 45 40 35 30 25 20(mg/mL) mM mM mM mM mM mM mM 60 654 638 623 607 592 576 560 55 613 597581 566 550 535 519 50 571 555 540 524 509 493 477 45 530 514 498 483467 452 436 40 488 472 457 441 426 410 394 35 447 431 415 400 384 369353 30 405 389 374 358 343 327 311

The results show that 1-DNJ-HCl concentration of 30 mg/mL (resulting infree 1-DNJ concentration of 25 mg/mL) in 50 mM Sodium Citrate bufferwill result in osmolality of 405 mOsm/kg. Ideally osmolality of theformulation for parenteral formulation would be 280-320 mOsm/kg so thatthe formulation would be isotonic with the target tissue or blood.Osmolality of about 500 mOsm/kg or below is generally regarded asacceptable for clinical trials as it does not cause deleterious healtheffects when administered to a human, such as irritation.

Example 8. Stability of 1-DNJ-HCl Formulation at Different Temperatures(Appearance and Color)

1-DNJ-HCl was dissolved in sodium citrate buffer. 1-DNJ-HClconcentration was 30 mg/mL (resulting free 1-DNJ concentration was 25mg/mL); sodium citrate buffer concentration was 50 mM at pH 5.0. Twobatches were made each containing about a thousand samples. L-12-026batch was stored for 6 months and E-13-014 batch was stored for 3months. The samples were stored under the nitrogen atmosphere(essentially 100% nitrogen). 1-DNJ-HCl solution in water was used ascontrol. The samples were stored at three different temperatures: 5°C.±3° C., 25° C.±2° C. or 40° C.±2° C. At 1, 3 and 6 months samples fromeach batch were collected and product stability was measured. Appearanceand color was measured in APHA units. Appearance of below 100 APHA unitsis generally regarded as acceptable for clinical trials as an indicationof stability.

Results:

Results are presented below in Tables 6.1-6.3. All samples appear asclear colorless solutions.

TABLE 6.1 Drug Product Stability Data for Appearance and Color at 5° C.± 3° C. Storage Conditions. Time (Months) Batch 0 1 3 6 L-12-026Conforms Conforms Conforms Conforms 0 0 0 0 E-13-014 Conforms ConformsConforms — 0 0 0

TABLE 6.2 Drug Product Stability Data for Appearance and Color at 25° C.± 2° C. Storage Conditions. Time (Months) Batch 0 1 3 6 L-12-026Conforms Conforms Conforms Conforms 0 0 <10 0 E-13-014 Conforms ConformsConforms — 0 0 0

TABLE 6.3 Drug Product Stability Data for Appearance and Color at 40° C.± 2° C. Storage Conditions. Time (Months) Batch 0 1 3 6 L-12-026Conforms Conforms Conforms Conforms 0 0 <10 10 E-13-014 ConformsConforms Conforms — 0 0 0

Results show that 1-DNJ-HCl formulation remains surprisingly stable. Nodiscoloration is observed during 6 months storage at temperaturesranging from about 2° C. to about 42° C.

Example 9. Stability of 1-DNJ-HCl Formulation at Different Temperatures(Osmolality)

1-DNJ-HCl was dissolved in sodium citrate buffer, under conditionsdescribed in Example 8. The samples were stored at two differenttemperatures: 5° C.±3° C. or 25° C.±2° C. At 1, 3 and 6 months samplesfrom each batch were collected and osmomolality was measured. InitialOsmomolality of the sample was about 405 mOsm/kg. Osmolality of about300-500 mOsm/kg is generally regarded as acceptable for clinical trials.

Results:

Results are presented below in Tables 7.1-7.2.

TABLE 7.1 Drug Product Stability Data for Osmolality at 5° C. ± 3° C.Storage Conditions. Time (Months) Batch 0 1 3 6 L-12-026 403 403 403 404E-13-014 403 405 405 —

TABLE 7.2 Drug Product Stability Data for Osmolality at 25° C. ± 2° C.Storage Conditions. Time (Months) Batch 0 1 3 6 E-13-014 403 407 407 —

Results show that the formulation demonstrates a remarkable stability.Osmolality remains unchanged after storage for 3 and 6 months attemperatures ranging from about 2° C. to about 27° C. Furthermore, notrend to decrease osmolality is observed. Without being bound by aparticular theory, the data indicates that osmolality is not going tosignificantly change during storage periods of about a year or longer.

Example 10. Stability of 1-DNJ-HCl Formulation at Different Temperatures(Particulate Matter)

1-DNJ-HCl was dissolved in sodium citrate buffer, under conditionsdescribed in Example 8. The samples were stored at two differenttemperatures: 5° C.±3° C. or 25° C.±2° C. At 1, 3 and 6 months samplesfrom each batch were collected and presense of particulate matter wasmeasured as number of particles per container. Particles of two sizeswere considered separately: particles greater or equal to 10 μm andparticles greater or equal to 25 μm. Presense of particules greater orequal to 10 μm in the amount of less or equal to about 6000 per 10 mLcontainter is generally regarded as acceptable for clinical trials.Presense of particules greater or equal to 25 μm in the amount of lessor equal to about 600 per 10 mL container is generally regarded asacceptable for clinical trials.

Results:

Results are presented below in Tables 8.1-8.2. Number of particles ineach sample is shown,

TABLE 8.1 Drug Product Stability Data for Particulate Matter at 5° C. ±3° C. Storage Conditions. Time (Months) Batch 0 1 3 6 L-12-026 ≥10 μm: 57 12 181 L-12-026 ≥25 μm: <1 3 0 9 E-13-014 ≥10 μm: 181 5 7 — E-13-014≥25 μm: 9 1 1

TABLE 8.2 Drug Product Stability Data for Particulate Matter at 25° C. ±2° C. Storage Conditions. Time (Months) Batch 0 1 3 6 E-13-014 ≥10 μm:181 1 3 — E-13-014 ≥25 μm: 9 0 1 —

Results show that the formulation demonstrates a remarkable stability.There is no increase of particular matter of any kind after storage for3 and 6 months at temperatures ranging from about 2° C. to about 27° C.Furthermore, increase in particulate matter greater or equal to 25 μm.It indicates that particles do not aggregate to form larger particles.Without being bound by a particular theory the data indicates thatamount of particles is not going to increase during storage periods ofabout a year or longer.

Example 11. Stability of 1-DNJ-HCl Formulation at Different Temperatures(pH)

1-DNJ-HCl was dissolved in sodium citrate buffer, under conditionsdescribed in Example 8. The samples were stored at three differenttemperatures: 5° C.±3° C., 25° C.±2° C. or 40° C.±2° C. At 1, 3 and 6months samples from each batch were collected and pH was measured.Initial pH of the solution was 5.0. Maintaining pH in the range of about4.8 to about 5.4 is considered as acceptable indication of stability.

Results:

Results are presented below in Tables 9.1-9.3.

TABLE 9.1 Drug Product Stability Data for pH at 5° C. ± 3° C. StorageConditions. Time (Months) Batch 0 1 3 6 L-12-026 5.1 5.0 5.0 5.0E-13-014 4.9 4.9 5.0 —

TABLE 9.2 Drug Product Stability Data for pH at 25° C. ± 2° C. StorageConditions. Time (Months) Batch 0 1 3 6 L-12-026 5.1 5.0 5.0 5.0E-13-014 4.9 5.0 5.0 —

TABLE 9.3 Drug Product Stability Data for Appearance and Color at 40° C.± 2° C. Storage Conditions. Time (Months) Batch 0 1 3 6 L-12-026 5.1 5.05.0 5.0 E-13-014 4.9 4.9 5.0 —

Results show that the formulation demonstrates remarkable stability.There is no change of any kind in pH after storage for 3 and 6 months attemperatures ranging from about 2° C. to about 42° C. Without beingbound by a particular theory the data indicates that pH of theformulation is not going to increase during storage periods of about ayear or longer.

Example 12. Stability of 1-DNJ-HCl Formulation at Different Temperatures(Concentration)

1-DNJ-HCl was dissolved in sodium citrate buffer, under conditionsdescribed in Example 8. The samples were stored at three differenttemperatures: 5° C.±3° C., 25° C.±2° C. or 40° C.±2° C., At 1, 3 and 6months samples from each batch were collected and concentration of 1-DNJas a free base was measured. Initial concentration of 1-DNJ as a freebase in the solution was about 25 mg/mL (corresponding to 30 mg/mL of1-DNJ HCl). Maintaining concentration of 1-DNJ as a free base in therange of about 22 to about 27 mg/mL (90-110%) is considered asacceptable indication of stability.

Results:

Results are presented below in Tables 10.1-10.3.

TABLE 10.1 Drug Product Stability Data for pH at 5° C. ± 3° C. StorageConditions. Time (Months) Batch 0 1 3 6 L-12-026 26.9 (110%) 25.5 (104%)25.6 (104%) 26.0 (106%) E-13-014 25.7 (103%) 26.1 (104%) 27.0 (108%) —

TABLE 10.2 Drug Product Stability Data for pH at 25° C. ± 2° C. StorageConditions. Time (Months) Batch 0 1 3 6 L-12-026 26.9 (110%) 25.7 (105%)25.6 (104%) 25.9 (106%) E-13-014 25.7 (103%) 25.2 (101%) 25.7 (103%) —

TABLE 10.3 Drug Product Stability Data for Appearance and Color at 40°C. ± 2° C. Storage Conditions. Time (Months) Batch 0 1 3 6 L-12-026 26.9(110%) 25.5 (104%) 24.4 (99%)  26.2 (107%) E-13-014 25.7 (103%) 25.6(102%) 25.1 (100%) —

Results show that the formulation demonstrates a remarkable stability.There is no significant change of in concentration after storage for 3and 6 months at temperatures ranging from about 2° C. to about 42° C.High concentration value at initial time point was caused by variabilityof reference standard injections and was not determined to beformulation or stability related. There is no visible pattern forincrease or decrease in concentration. Without being bound by aparticular theory the data indicates that concentration of theformulation is not going to increase during storage periods of about ayear or longer.

Example 13. Hemolytic Potential of 1-DNJ-HCl Formulation in Human, Rat,and Cynomolgus Monkey Whole Blood

1-DNJ-HCl was dissolved in sodium citrate buffer. 1-DNJ-HClconcentration was 30 mg/mL (resulting free 1-DNJ concentration was 25mg/mL); sodium citrate buffer concentration was 50 mM at pH 5.0.1-DNJ-HCl formulation was added to whole blood samples of male human,rat, and cynomolgus monkey. 500 μM of 1-DNJ-HCl formulation was mixedwith 500 μM of blood sample. Buffer served as a negative control, whileTriton-X 100 (Lysing agent) served as a positive control. Additionallyblood sample without any additions was used as a negative control.

The samples were stored at 4° C. Blood samples were incubated at 37° C.for 30 minutes with the 1-DNJ-HCl formulation, after which the sampleswere centrifuged. The amount of hemoglobin in the supernatant from eachcentrifuged sample was measured using a commercially availablehemoglobin assay kit (BioAssay Systems, QuantiChrom™ Hemoglobin AssayKit) and the Tecan microplate reader.

Results:

Results are presented below in Table 11.

TABLE 11 Hemolytic Potential of 1-DNJ-HCl in Whole Blood Samples: 500 μLblood volume was mixed with 500 μL % of the Avg OD Std Dev HemoglobinRBC Species following: Dilution (n = 4) (mg/dL) (mg/dL) lysis Human 30mg/mL 1 0.0755 0.005 19 0.0% AT2220 Vehicle 1 0.0706 0.005 18 0.0% — 10.2568 0.030 64 0.7% 0.1% 100 0.2768 0.002 6899 100.0% Triton X-100Monkey 30 mg/mL 1 0.0649 0.004 16 0.1% AT2220 Vehicle 1 0.0488 0.001 120.0% — 1 0.2989 0.036 74 1.3% 0.1% 100 0.2000 0.004 4984 100.0% TritonX-100 Rat 30 mg/mL 1 0.3535 0.078 88 −0.3% AT2220 Vehicle 1 0.4304 0.041107 0.0% — 1 0.5634 0.021 140 0.5% 0.1% 100 0.2567 0.007 6398 100.0%Triton X-100

Results show that the formulation demonstrates a surprising lack ofhemolytic potential. No hemolysis of erythrocytes in human, SpragueDawley rat, or Cynomolgus monkey whole blood was detected with thevehicle or undiluted 1-DNJ-HCl formulation mixtures. The 1-DNJ-HClformulation and its vehicle did not result in any detectable hemolysisin whole blood from any of the three species tested in this assay.Without being bound by a particular theory the data indicates that theformulation is not going to cause deleterious health effects such aspain or irritation when administered to the subject. The 1-DNJ-HClformulation demonstrates a remarkable safety as it does not causevascular irritation.

***

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, that while the invention hereinhas been described with reference to particular embodiments, it is to beunderstood that these embodiments are merely illustrative of theprinciples and applications of the present invention. It is therefore tobe understood that numerous modifications may be made to theillustrative embodiments and that other arrangements may be devisedwithout departing from the spirit and scope of the present invention asdefined by the appended claims.

In the claims articles such as “a,” “an,” and “the” may mean one or morethan one unless indicated to the contrary or otherwise evident from thecontext. Claims or descriptions that include “or” between one or moremembers of a group are considered satisfied if one, more than one, orall of the group members are present in, employed in, or otherwiserelevant to a given product or process unless indicated to the contraryor otherwise evident from the context. The invention includesembodiments in which exactly one member of the group is present in,employed in, or otherwise relevant to a given product or process. Theinvention includes embodiments in which more than one, or all of thegroup members are present in, employed in, or otherwise relevant to agiven product or process. Furthermore, it is to be understood that theinvention encompasses all variations, combinations, and permutations inwhich one or more limitations, elements, clauses, descriptive terms,etc., from one or more of the listed claims is introduced into anotherclaim. For example, any claim that is dependent on another claim can bemodified to include one or more limitations found in any other claimthat is dependent on the same base claim.

Where elements are presented as lists, e.g., in Markush group format, itis to be understood that each subgroup of the elements is alsodisclosed, and any element(s) can be removed from the group. It shouldit be understood that, in general, where the invention, or aspects ofthe invention, is/are referred to as comprising particular elements,features, etc., certain embodiments of the invention or aspects of theinvention consist, or consist essentially of, such elements, features,etc. For purposes of simplicity those embodiments have not beenspecifically set forth in haec verba herein. It is noted that the term“comprising” is intended to be open and permits the inclusion ofadditional elements or steps.

Where ranges are given, endpoints are included. Furthermore, it is to beunderstood that unless otherwise indicated or otherwise evident from thecontext and understanding of one of ordinary skill in the art, valuesthat are expressed as ranges can assume any specific value or subrangewithin the stated ranges in different embodiments of the invention, tothe tenth of the unit of the lower limit of the range, unless thecontext clearly dictates otherwise.

In addition, it is to be understood that any particular embodiment ofthe present invention that falls within the prior art may be explicitlyexcluded from any one or more of the claims. Since such embodiments aredeemed to be known to one of ordinary skill in the art, they may beexcluded even if the exclusion is not set forth explicitly herein. Anyparticular embodiment of the compositions of the invention (e.g., anytargeting moiety, any disease, disorder, and/or condition, any linkingagent, any method of administration, any therapeutic application, etc.)can be excluded from any one or more claims, for any reason, whether ornot related to the existence of prior art.

Publications discussed above and throughout the text are provided solelyfor their disclosure prior to the filing date of the presentapplication. Nothing herein is to be construed as an admission that theinventors are not entitled to antedate such disclosure by virtue ofprior disclosure.

The invention claimed is:
 1. A pharmaceutical composition comprising: a)an active agent selected from 1-deoxynojirimycin, a pharmaceuticallyacceptable salt thereof, or a derivative thereof, and b) a buffer,wherein the active agent is present at a concentration of from about 1mg/mL to about 100 mg/mL.
 2. The pharmaceutical composition of claim 1,wherein the active agent comprises 1-deoxynojirimycin.
 3. Thepharmaceutical composition of claim 1 wherein the active agent comprises1-deoxynojirimycin hydrochloride.
 4. The pharmaceutical composition ofclaim 1, wherein the active agent comprises N-butyl-deoxynojirimycin, ora pharmaceutically acceptable salt thereof.
 5. The pharmaceuticalcomposition of claim 1, wherein the pharmaceutical composition isformulated for intravenous administration.
 6. The pharmaceuticalcomposition of claim 1, wherein the pharmaceutical composition is at apH of from about 4 to about
 6. 7. The pharmaceutical composition ofclaim 1, wherein the active agent is present at a concentration of fromabout 1 mg/mL to about 60 mg/mL.
 8. The pharmaceutical composition ofclaim 1, wherein the active agent is present at a concentration of fromabout 25 mg/mL to about 30 mg/mL.
 9. The pharmaceutical composition ofclaim 1, wherein the buffer is present at a concentration of about 20 mMto about 75 mM.
 10. The pharmaceutical composition of claim 1, whereinthe buffer is present at a concentration of about 40 mM to about 50 mM.11. The pharmaceutical composition of claim 1, wherein the buffercomprises a citrate buffer.
 12. The pharmaceutical composition of claim1, further comprising a chelating agent.
 13. The pharmaceuticalcomposition of claim 12, wherein the chelating agent comprises EDTA. 14.The pharmaceutical composition of claim 13, wherein the EDTA is presentat a concentration of about 0.005% to about 0.25% weight by volume. 15.The pharmaceutical composition of claim 1, wherein the pharmaceuticalcomposition is shelf-stable for at least about one year under anatmosphere selected from nitrogen, air or a combination thereof, and atemperature of from about 2° C. to about 42° C.
 16. A pharmaceuticalcomposition comprising a) an active agent comprising N-butyl-deoxynojirimycin, or a pharmaceutically acceptable salt thereof ,and b) a buffer, wherein the active agent is present at a concentrationof from about 1 mg/mL to about 100 mg/mL and the buffer is present at aconcentration of about 20 mM to about 75 mM.
 17. The pharmaceuticalcomposition of claim 16, wherein the buffer comprises a citrate buffer.18. A method for increasing the stability of a formulation comprising anactive agent selected from 1-deoxynojirimycin, a pharmaceuticallyacceptable salt thereof, or a derivative thereof, the method comprisingintroducing a buffer into the formulation, wherein the active agent ispresent at a concentration of from about 1 mg/ml to about 100 mg/mL. 19.The method of claim 18, wherein the active agent comprisesN-butyl-deoxynojirimycin, or a pharmaceutically acceptable salt thereof.20. The method of claim 18, wherein the buffer comprises a citratebuffer.